An improved protocol for DNA extraction from the faeces of giant panda

摘要

An improved method that facilitates the extraction of PCR-compatible faecal DNA from giant panda's faeces is described. The method involves a novel preprocessing step in DNA extraction. The faeces is washed two or three times with precooled acetone, which removes numerous potential PCR inhibitors, and then digested with proteinase K. The DNA is purified with phenol/chloroform. The faecal DNA obtained is sufficiently pure to support reliable amplification, and is applied as template DNA to amplify a portion of the giant panda brain derived neurotrophic factor ( BDNF) gene and mitochondrial cytochrome b gene. The sequenced results of PCR products confirm that the extracted DNA is from the giant panda. Comparison with the PCR products demonstrates that the faecal DNA extracted by the improved protocol is better than the faecal DNA extracted without acetone preprocessing. This protocol provides a reliable method for faecal DNA extraction and has potential significance for molecular genetic study of the giant panda [ Acta Zoologica Sinica 49(5): 670 - 674, 2003] .